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Cell Tracking in Multimodal Holographic Microscope Using Transmission Mode
Blašťáková, Lucia ; Křížová, Aneta (referee) ; Štrbková, Lenka (advisor)
This thesis is focused on the monitoring of cell dynamics in multimodal holographic microscope using transmission mode. The goal of this experiment is a quantitative description of the epithelial-mesenchymal transition of cancer cells NMuMG. The introduction briefly describes a history of interference microscopy and its applications, design of multimodal holographic microscope Q-Phase and work with the program used for data processing. Next follows a brief description of parameters that were selected to quantify the epithelial-mesenchymal transition. In conclusion, all results from the observation of epithelial-mesenchymal transition were summarized.
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Cell response to genotoxic stress-based anti-cancer therapies
Imrichová, Terezie ; Hodný, Zdeněk (advisor) ; Rossmeislová, Lenka (referee) ; Rotrekl, Vladimír (referee)
The dissertation deals with a cell response to genotoxic stress, specifically to anti-cancer treatments with a genotoxic mechanism of action. In principle, cells can respond to these perturbing stimuli in several ways: in case of severe DNA damage, they usually undergo apoptosis or enter senescence. In case of minor DNA damage, or upon defective checkpoint mechanisms, they may continue the cell cycle, either with successfully repaired DNA or with mutations of various kind. Thanks to selection pressure, the mutations that provide cells with a certain growth advantage under conditions of continuing genotoxic stress, gradually accumulate and render the tumor treatment-resistant. In my thesis, I focus on several aspects of this whole process. First, I participated in a characterization of a radioresistant and anoikis-resistant population of prostate cancer cells. This population was generated by irradiating cells 35 times by 2 Gy, a regime used in clinics. After this treatment, a population of low-adherent cells emerged that demonstrated increased expression of EMT- and stem cell markers. The low-adherent state of these cells was maintained by Snail signaling and their anoikis resistance by ERK1/2 signaling. Interestingly, after a protracted period of time, these cells were able to re-adhere and...
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Cell response to genotoxic stress-based anti-cancer therapies
Imrichová, Terezie ; Hodný, Zdeněk (advisor) ; Rossmeislová, Lenka (referee) ; Rotrekl, Vladimír (referee)
The dissertation deals with a cell response to genotoxic stress, specifically to anti-cancer treatments with a genotoxic mechanism of action. In principle, cells can respond to these perturbing stimuli in several ways: in case of severe DNA damage, they usually undergo apoptosis or enter senescence. In case of minor DNA damage, or upon defective checkpoint mechanisms, they may continue the cell cycle, either with successfully repaired DNA or with mutations of various kind. Thanks to selection pressure, the mutations that provide cells with a certain growth advantage under conditions of continuing genotoxic stress, gradually accumulate and render the tumor treatment-resistant. In my thesis, I focus on several aspects of this whole process. First, I participated in a characterization of a radioresistant and anoikis-resistant population of prostate cancer cells. This population was generated by irradiating cells 35 times by 2 Gy, a regime used in clinics. After this treatment, a population of low-adherent cells emerged that demonstrated increased expression of EMT- and stem cell markers. The low-adherent state of these cells was maintained by Snail signaling and their anoikis resistance by ERK1/2 signaling. Interestingly, after a protracted period of time, these cells were able to re-adhere and...
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Role of transcription factor Snail in mechanism of development of radiorezistence in prostate carcinoma cell lines
Davidová, Eliška ; Hodný, Zdeněk (advisor) ; Benada, Jan (referee)
The frequent cause of failure of prostate carcinoma radiotherapy and chemotherapy is the emergence of resistance and a progress into the essentially incurable metastatic form of disease. Although the mechanisms of the radioresistance and chemoresistance are still not well understood, recent studies indicate that transcription factor Snail, a key mediator of the epithelial-mesenchymal transition and subsequent metastasis formation, plays a critical role in the development of the chemoresistance and radioresistance in the tumor cells. As the activation of the optimal DNA damage response pathway is the determining factor for the cell survival after chemotherapy and radiotherapy, we hypothesized the role of Snail in the transcription regulation of these processes. In this study, we first analyzed the relationship between Snail and ATM kinase, as the ATM was recently reported to regulate stability of Snail by its phosphorylation. Although, we observed a modest effect of ATM inhibition on Snail levels after cancer cells exposure to ionizing radiation, we did not fully reproduced the recently published findings. Furthermore, we evaluated the role of Snail in transcription regulation of cyclin-dependent kinase inhibitor p21waf1/cip1 . Our data point towards the suppressive role of Snail in p21waf1/cip1...
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Cell Tracking in Multimodal Holographic Microscope Using Transmission Mode
Blašťáková, Lucia ; Křížová, Aneta (referee) ; Štrbková, Lenka (advisor)
This thesis is focused on the monitoring of cell dynamics in multimodal holographic microscope using transmission mode. The goal of this experiment is a quantitative description of the epithelial-mesenchymal transition of cancer cells NMuMG. The introduction briefly describes a history of interference microscopy and its applications, design of multimodal holographic microscope Q-Phase and work with the program used for data processing. Next follows a brief description of parameters that were selected to quantify the epithelial-mesenchymal transition. In conclusion, all results from the observation of epithelial-mesenchymal transition were summarized.
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